Early Evolutionary History of Dinoflagellates and Apicomplexans (alveolata) as Inferred from Hsp90 and Actin Phylogenies

Three extremely diverse groups of unicellular eukaryotes comprise the Alveolata: ciliates, dinoflagellates, and apicomplexans. The vast phenotypic distances between the three groups along with the enigmatic distribution of plastids and the economic and medical importance of several representative species (e.g. Plasmodium, Toxoplasma, Perkinsus, and Pfiesteria) have stimulated a great deal of speculation on the early evolutionary history of alveolates. A robust phylogenetic framework for alveolate diversity will provide the context necessary for understanding the basic biological properties of the group and for developing appropriate strategies for management. We addressed the earliest stages of alveolate evolution by sequencing heat shock protein 90 (hsp90) genes from several ciliates, apicomplexans, and dinoflagellates, including key species thought to represent early diverging lineages: Oxyrrhis marina, Perkinsus marinus, Cryptosporidium parvum, and the eugregarine Monocystis agilis. Moreover, by sequencing the actin gene from Monocystis, we were able to examine the sister relationship between gregarines and cryptosporidians with a three-protein concatenated data set (hsp90, actin, and b-tubulin). Phylogenetic analyses of the hsp90 data set provided a robust topology for alveolate relationships: Alveolates were monophyletic and apicomplexans and dinoflagellates formed sister groups to the exclusion of ciliates. Oxyrrhis formed the earliest diverging sister lineage to the ‘‘core’’ dinoflagellates, and Perkinsus formed the earliest diverging sister lineage to the Oxyrrhis–dinoflagellate clade. This topology was strongly supported in all analyses and by a unique indel shared by Oxyrrhis and dinoflagellates. A sister relationship between Cryptosporidium andMonocystis was weakly supported by the hsp90 data set but strongly supported by the three-protein concatenated data set.